RNA-seq of mouse wildtype HSCs purified from recipient mice co-transplanted with wildtype or different types of lymphoid cell deficient HSCs
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ABSTRACT: To investigate how HSCs functionally compensate for the B cell deficiencies of other HSCs within an organism, we co-transplanted wildtype (WT) HSCs and lineage-deficient HSCs into lethally irradiated WT recipient mice. WT HSCs were then purified from the bone marrow of recipient mice for RNA isolation and sequencing. The purpose is to determine whether there are common genes shared between compensating WT HSCs in the WT co-transplanted with uMT-/- (B6.129S2(B6)-Ighmtm1Cgn/J) and WT co-transplanted with NSG (NOD-scid IL2Rgamma null).
Project description:Murine long-term hematopoietic stem cells (HSCs), short-term HSCs and multipotent progenitor cells (MPPs) were isolated from bone marrow and expression profiled on Affy chips. The behavior of maternal-specific imprinting genes, particularly in the H19-Igf2 locus, was focused on, to see if any might be involved in maintaining quiescence of long-term stem cells.
Project description:The aim of the study was to analyse gene expression differences in LT and ST HSCs in BM of TEL-SYK transplanted versus control C57BL/6mice
Project description:This SuperSeries is composed of the following subset Series: GSE35110: Gene expression profiling in WT and Hif1a-/- HSCs GSE35111: Gene expression profiling in BCR/ABL expressing LSCs and BCR/ABL expressing Hif1a-/-LSCs Refer to individual Series
Project description:Gene expression of HSC (Lin-Sca-Kit+Flt3-) hematopoietic stem cells from Dicer D/+ mice was compared with LT-HSC from DicerD/D mice. BM cells were noncompetitively transplanted into irradiant recipients, and deletion induced by poly(IC) injection. Donor-derived HSCs were isolated by FACS sorting of recipient bone marrow. Biological replicates of each (2 per genotype) were generated and expression profiles were determined by hybridization to Affymetrix Moe430_2 arrays.
Project description:Murine WBP1L-deficient (Wbp1l-/-) hematopoietic stem and progenitor cells engraft significantly better than wild-type (WT) cells in competitive transplantation assays. To analyze the mechanism, Wbp1l-/- or WT bone marrow cells (both Ly5.2+Ly5.1-) were each mixed 1:1 with WT competitor cells (Ly5.2+Ly5.1+) and the mixture was transplanted into the lethally irradiated recipient mice. 22 weeks after the transplantation the donor Ly5.2+Ly5.1- LSK cells (Wbp1l-/- or WT) were purified from the bone marrow of recipient mice, RNA was extracted and subjected to RNA sequencing.
Project description:WT bone marrow (BM) was transplanted into WT or CMO recipient mice. For 12 weeks the WT cells (including WT hematopoietic stem cells; HSCs) were exposed to WT or CMO BM niche. After 12 weeks, the BM was isolated, the WT donor HSCs were sorted, RNA isolated and RNA sequencing was performed. This experiment allowed us to investigate the effect of the WT or CMO BM niche on WT HSCs.
Project description:To test the long-term effects of lack of Dnmt3a on mouse HSCs, we established a cohort of lethally irradiated mice transplanted with Dnmt3a-KO or WT HSC control cells. Dnmt3a deletion in purified HSCs transplanted alone leads to an array of hematologic disorders that models the spectrum of disorders seen in human malignancies. We investigated DNA methylation alterations in disease and control mice by RRBS. We generated an average of 23.81 million reads per sample (18.97 - 31.67 million) of which an average of 18.07 million could be mapped to the mm9 genome (2.78 - 26.42 million). We achieved an average CpG coverage depth of 31.79-fold (8.09 - 47.44-fold) and average bisulfite conversion efficiency of 99.91% (99.87 - 99.95%). Reduced representation bisulfite sequencing of cells using Illumina HiSeq 2000 and 2500
Project description:We performed single cell whole transcriptome (Smart-seq3) analysis of hematopoietic stem and progenitor cells long-term replenished by transplanted single Vwf-tdTomato+ P-HSCs (2290 total cells from 7 reconstituted mice) and Vwf-tdTomato Multi-HSCs (2478 total cells from 8 reconstituted mice), as well as control cells from normal mouse bone marrow.
Project description:Hematopoietic stem cells (HSCs) can regenerate the entire hematopoietic system in vivo, providing the most relevant criteria to measure candidate HSCs derived from human embryonic stem cell (hESC) or induced pluripotent stem cell (hiPSC) sources. Here, we show that unlike primitive hematopoietic cells derived from hESCs, phenotypically identical cells derived from hiPSC are more permissive to graft the bone marrow of xenotransplantation recipients. Despite establishment of bone marrow graft, hiPSC-derived cells fail to demonstrate hematopoietic differentiation in vivo. However, once removed from recipient bone marrow, hiPSC-derived grafts were capable of in vitro multilineage hematopoietic differentiation, indicating that xenograft imparts a restriction to in vivo hematopoietic progression. This failure to regenerate multilineage hematopoiesis in vivo was attributed to the inability to downregulate key microRNAs involved in hematopoiesis. Based on these analyses, our study indicates that hiPSCs provide a beneficial source of pluripotent stem cell-derived hematopoietic cells for transplantation compared with hESCs. Since use of the human-mouse xenograft models prevents detection of putative hiPSC-derived HSCs, we suggest that new preclinical models should be explored to fully evaluate cells generated from hiPSC sources. Human pluripotent stem cell-derived hematopoietic cells were isolated and qPCR-based microRNA profiling was performed.
Project description:Analysis of highly purified long-term hematopoietic stem cells (LT-HSCs) irradiated at 0Gy, 0.02Gy, 0.1Gy and 0.5Gy six months after transplantation. Results provide insight into the molecular mechanisms underlying multiple aspects of LT-HSCs premature ageing after low doses of γ-irradiation (0.02Gy). Four samples were analyzed and correlated with the control group (0Gy).